Bottom Line:
In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization.The latter was performed using both confocal and transmission electron microscopy.We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation.

ABSTRACTBoron nitride nanotubes (BNNTs) have generated considerable interest within the scientific community by virtue of their unique physical properties, which can be exploited in the biomedical field. In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization. The latter was performed using both confocal and transmission electron microscopy. Finally, we investigated myoblast differentiation in the presence of BNNTs, evaluating the protein synthesis of differentiating cells, myotube formation, and expression of some constitutive myoblastic markers, such as MyoD and Cx43, by reverse transcription - polymerase chain reaction and Western blot analysis. We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation.

f8-ijn-5-285: RT-PCR results for MyoD and Cx43 expression in C2C12 cells differentiated in presence of 0, 5, and 10 μg/ml of PLL-BNNTs; GAPDH was used as internal standard (a) Western blot results for MyoD and Cx43 production in the same cultures; β-actin was used as internal standard (b) Immunocytochemical analysis for MyoD expression on BNNT-treated cell samples (c) and controls without BNNTs (d) “Bn” indicates visible dotting in the cytoplasm due to BNNTs, the arrows indicate MyoD positive cells at nuclear level, while the arrowhead indicates MyoD positive cells at cytoplasm level.Abbreviations: BNNTs, boron nitride nanotubes; H&E, hematoxylin–eosin; PLL, poly-l-lysine; RT-PCR, reverse transcription – polymerase chain reaction.

Mentions:
Figure 8a shows MyoD and Cx43 mRNA levels in differentiated C2C12 cells treated with 0, 5 and 10 μg/mL of PLL-BNNTs, as obtained by RT-PCR. These results highlighted that the expression of both Cx43 and MyoD genes was not drastically affected by the administration of BNNTs at a concentration lower than 10 μg/mL: Cx43 and MyoD mRNAs were found to be well expressed in all the tested samples.

f8-ijn-5-285: RT-PCR results for MyoD and Cx43 expression in C2C12 cells differentiated in presence of 0, 5, and 10 μg/ml of PLL-BNNTs; GAPDH was used as internal standard (a) Western blot results for MyoD and Cx43 production in the same cultures; β-actin was used as internal standard (b) Immunocytochemical analysis for MyoD expression on BNNT-treated cell samples (c) and controls without BNNTs (d) “Bn” indicates visible dotting in the cytoplasm due to BNNTs, the arrows indicate MyoD positive cells at nuclear level, while the arrowhead indicates MyoD positive cells at cytoplasm level.Abbreviations: BNNTs, boron nitride nanotubes; H&E, hematoxylin–eosin; PLL, poly-l-lysine; RT-PCR, reverse transcription – polymerase chain reaction.

Mentions:
Figure 8a shows MyoD and Cx43 mRNA levels in differentiated C2C12 cells treated with 0, 5 and 10 μg/mL of PLL-BNNTs, as obtained by RT-PCR. These results highlighted that the expression of both Cx43 and MyoD genes was not drastically affected by the administration of BNNTs at a concentration lower than 10 μg/mL: Cx43 and MyoD mRNAs were found to be well expressed in all the tested samples.

Bottom Line:
In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization.The latter was performed using both confocal and transmission electron microscopy.We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation.

ABSTRACTBoron nitride nanotubes (BNNTs) have generated considerable interest within the scientific community by virtue of their unique physical properties, which can be exploited in the biomedical field. In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization. The latter was performed using both confocal and transmission electron microscopy. Finally, we investigated myoblast differentiation in the presence of BNNTs, evaluating the protein synthesis of differentiating cells, myotube formation, and expression of some constitutive myoblastic markers, such as MyoD and Cx43, by reverse transcription - polymerase chain reaction and Western blot analysis. We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation.